The preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation.

Content Provider	Europe PMC
Author	Kroeger, Kelly Collins, Michelle Ugozzoli, Luis
Copyright Year	2009
Abstract	It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer were specifically developed to transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells.This video demonstrates how to establish primary hematopoietic cell cultures from murine bone marrow, and then prepare them for electroporation in the MXcell system. We begin by isolating femur and tibia. Bone marrow from both femur and tibia are then harvested and cultures are established. Cultured bone marrow cells are then transfected and analyzed.
Journal	Journal of Visualized Experiments : JoVE
PubMed Central reference number	PMC2763291
Issue Number	23
PubMed reference number	19229174
e-ISSN	1940087X
DOI	10.3791/1026
Language	English
Publisher	MyJove Corporation
Publisher Date	2009-01-06
Access Restriction	Open
Rights License	This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Copyright © 2009, Journal of Visualized Experiments
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Chemical Engineering Neuroscience Biochemistry, Genetics and Molecular Biology